Determination Of Ursolic Acid Research Articles

Determination of ursolic acid in ethanolic extract of Rotheca serrata (L.) Steane & Mabb. by HPLC method

Rotheca serrata (L.) Steane & Mabb. a plant having significant medicinal value is used in treating a fever, rheumatism, inflammation, pain, and malarial fever. Ursolic acid has a five-ringed pentacyclic structure belongs to the family of triterpenoids. It has antioxidant, antitumor, and hepato-protective properties. The method in HPLC was developed on a SHIMADZU, Shimpack ODS (25 cm × 4.6 mm, 5 µm) column. The mobile phase consisted of a 70:25:5 (v/v/v) mixture of methanol, acetonitrile, and 0.02 M ammonium acetate buffer (pH 3.5). A flow rate of 1.2 mL/min and λmax of 215 nm. Ursolic acid from the extract and standard was eluted at the same retention time of 4.1 min. 20–120 µg/mL was the linearity range chosen and was found to be linear (R 2=0.996). The value of LOD was 0.85 µg/mL and LOQ was 1.96 µg/mL. The method for analysis of ursolic was found to be accurate with good repeatability.

MEASUREMENT UNCERTAINTY EVALUATION OF ANALYTICAL METHOD FOR QUANTITATIVE DETERMINATION OF URSOLIC AND OLEANOLIC ACIDS IN APPLE PROCESSING AGROINDUSTRIAL WASTE MATERIAL

Background: Apple pomace represents a low-cost and rich source of bioactive compounds with valuable properties - ursolic acid (UA) and oleanolic acid (OA). Due to the wide range of applications in pharmaceutical and nutraceutical industries, these compounds have a high commercial value, and possessing a suitable analytical method with measurement uncertainty is of great significance and practicability. Aim: The purpose of the present work was to estimate detailed measurement uncertainty for the validated HPLC method combined with the extraction procedure for the determination of UA and OA in apple pomace. Methods: The chromatographic analysis using LC-20AD Prominence Shimadzu System and ultrasound-assisted extraction using the ultrasonic bath DW-5200DTS were performed to obtain and determine UA and OA in apple pomace. The process of measurement uncertainty evaluation was performed by the Ishikawa diagram and a combination of bottom-up and top-down approaches. Results: The content of UA and OA (mg/g) in apple pomace with the value of the expanded uncertainty was calculated, which is 7.06 ± 0.647 mg/g (k=1.96; P=95%) and 4.70 ± 0.422 mg/g (k=1.96; P=95%), respectively. Six sources of all the contributors of uncertainties were observed that affected the measurement. Discussion: The A-type standard uncertainty value was 3 times less than the B-type standard uncertainty for both analytes. The results show that B-type standard uncertainty is a major contributor, and the value of the expanded uncertainty of the validated method will not change from test to test in the same laboratory conditions. Conclusions: The methodology described in this work explains well the details and practical aspects of the hybrid approach using the method validation data and proposes step-by-step instructions to evaluate the measurement uncertainty of the quantitative method.

Determination of Ursolic Acid in Extracts From Ligustri lucidum Fruit Using an Electrochemical Method.

In this work, we reported a facile wet chemical method for depositing Pt nanoparticles on the surface of boron nitride nanosheets (BNNS-Pt NPs). The deposited nanocomposite was applied for glassy carbon electrode surface modification. The modified electrode was then used for detecting ursolic acid (UA). The results indicate that the BNNS-Pt NPs exhibited excellent electrocatalytic activity toward UA oxidation compared with that of the bare glassy carbon electrode (GCE) and Pt NPs/GCE. The UA oxidation currents is linearly related its concentration from 1 to 1,200 pM. The limit of detection can be calculated to be 0.5 pM. In addition, the UA sensor was also successfully used for the determination of UA in Ligustri lucidum fruit samples.

A metal organic framework polymer monolithic column as a novel adsorbent for on-line solid phase extraction and determination of ursolic acid in Chinese herbal medicine

A metal organic framework (MOF)-polymer monolithic column was prepared by redox initiation using modified MOF and N-methylolacrylamide (NMA) as co-monomers. The obtained monolithic column was characterized by scanning electron microscopy (SEM) and nitrogen adsorption-desorption isotherm measurement. It was used as a solid phase extraction (SPE) absorbent for the online enrichment of ursolic acid (UA) by high performance liquid chromatography. The adsorption amount of UA on the monolith was compared with that of silica gel-C18 adsorbent and the monolith without MOF material. The MOF-polymer monolithic column showed high selectivity and good permeability. Under the optimum conditions for extraction and determination, the calibration equation was y = 79.854× + 0.1939; the linear range was 0.001–0.9 mg/mL; the linear regression coefficient was 0.9993; the limit of detection (LOD) and the limit of quantification (LOQ) were 0.17 μg/mL and 0.57 μg/mL, respectively; the inter-day and intra-day accuracies were <6.44%; the recovery was in the range of 86.52–105.26%. The MOF-polymer monolithic column was successfully used as SPE column for enrichment and determination of UA in Chinese herbal medicine.

Electrochemical detection of ursolic acid from spruce (Picea abies) essential oils using modified amperometric microsensors

Ursolic acid (UA) is a natural pentacyclic triterpenoid carboxylic acid found in some medicinal plant species. In this paper, amperometric microsensors based on a powder which contained graphite (G) and carbon nanoparticles (CN) (G-CN) unmodified and modified with chitosan (CHIT) and tetraphenyl-porphine cobalt(II) (Co(II)TPP) are proposed for the analysis of UA plant source essential oils obtained from spruce (Picea Abies). Differential pulse voltammetry (DPV) was used to optimize the method and for the determination of ursolic acid from different types of botanical samples. The optimum working pH was 5.00 for the G-CN and (Co(II)TPP)/G-CN microsensors and pH 3.00 for CHIT/G-CN in the presence of a 0.1 mol L−1 KCl supporting electrolyte. The linear concentration ranges for ursolic acid (UA) were between 0.1 and 100 µmol L−1 for the unmodified microsensor (G-CN), 0.01, 1 µmol L−1 for the microsensor modified with chitosan (CHIT/G-CN), and 0.01 and 10 µmol L−1 for the microsensor modified with (Co(II)TPP)/G-CN). It is the first time these amperometric microsensors have been used for the reliable analysis of ursolic acid (UA) in three original botanical samples obtained from different parts of spruce (Picea abies): resin essential oil, cons essential oil, cons and sprouts essential oil, with recovery rate values up to 99.29%.

HPLC determination of Ursolic acid in flowers of an Iranian Pomegranate (Punica granatum L.) Cultivar

Ursolic acid (UA), is a major bioactive compound in several traditional medicinal plants including pomegranate (Punica granatum L.) flower. This study presents the HPLC analysis of UA content in flowers of an Iranian pomegranate cultivar. The analytical method’s accuracy and the repeatability were satisfactory. The Linear ranges were 100-400 µg/ml for UA with a good correlation. On the average, the recovery rate of UA was 100%. Here, the extraction of UA from an Iranian pomegranate flower cultivar by employing an ultrasound-assisted extraction method was reported, and several extraction solvents were studied to improve extraction yield of UA. Remarkable quantities of UA were found in cultivar of pomegranate flower evaluated. Our results showed that Iranian pomegranate flower cv. Ghorj Tafti Torsh is a good source for extraction and isolation of UA.&#x0D;

Simultaneous Analysis of Ursolic Acid and Oleanolic Acid in Guava Leaves Using QuEChERS-Based Extraction Followed by High-Performance Liquid Chromatography.

In this paper, a novel method of QuEChERS-based extraction coupled with high-performance liquid chromatography has been developed for the simultaneous determination of ursolic acid (UA) and oleanolic acid (OA) in guava leaves. The QuEChERS-based extraction parameters, including the amount of added salt, vortex-assisted extraction time, and absorbent amount, and the chromatographic conditions were investigated for the analysis of UA and OA in guava leaves. Under the optimized conditions, the method showed good linearity over a range of 1–320 μg mL−1, with correlation coefficients above 0.999. The limits of detection of UA and OA were 0.18 and 0.36 μg mL−1, respectively. The intraday and interday precision were below 1.95 and 2.55%, respectively. The accuracies of the UA and OA determinations ranged from 97.4 to 111.4%. The contents of UA and OA in the guava leaf samples were 2.50 and 0.73 mg g−1, respectively. These results demonstrate that the developed method is applicable to the simultaneous determination of UA and OA in guava leaves.

A Simple and Reliable HPTLC Method for Determination of Ursolic Acid in Various Ocimum Species

A simple and reliable HPTLC method has been developed for the determination of Ursolic acid in various Ocimum species Viz. O. gratissium, O. sanctum and O. baslicum. The chromatographic separation was achieved in silica gel 60F254 HPTLC plate with Chlorform:Methanol (95:5 v/v) as a mobile phase. The quantification was performed in reflection/absorbance mode at a wavelength at 680 nm after post derivatization with vanillin-sulphuric acid reagent. The method showed good linear relationship (r2=0.983) over concentration range 2-10 µg/band. The LOD and LOQ were 1.76 µg/band and 6.02 µg/band for ursolic acid, respectively. The good results were achieved with respect precision (% RSD <2) and average recovery (97.78 %). The method was validated for its linearity, accuracy, precision, LOQ and LOD. The validated method was simple and reproducible can be used in overall quality control of Ocimum species and herbal formulation containing these species.

Determination of ursolic acid in fractionated leaf extracts of Ocimum gratissimum Linn and in developed herbal hepatoprotective tablet by HPTLC

BackgroundUrsolic acid was determined in dichloromethane and ethyl acetate fractions of methanolic extract of Ocimum gratissimum and in developed herbal hepatoprotective tablet by HPTLC method. MethodsHepatoprotective polyherbal formulation was prepared using five fractions of three plant extracts namely O. gratissimum, Butea monosperma and Bauhinia variegata. Among these three plants O. gratissimum contains ursolic acid. Chromatographic separation was performed on silica gel HPTLC plates with petroleum ether:ethyl acetate:acetone (8.2:1.8:0.1, v/v/v) as mobile phase. After drying, the plates were sprayed with 10% (v/v) ethanolic solution of sulfuric acid and heated to 120 °C for 3 min. Quantification was performed in absorbance/transmittance mode at a wavelength of 530 nm using a computer-controlled densitometer. ResultsThe presented method was validated for linearity 400–1200 (ng/spot), intraday precision % C.V. (0.58–1.97), and interday precision % C.V. (1.46–2.22). Correlation coefficient (r2 = 0.9960), detection limits as well as recovery values (97.5%–98.22%) were found to be satisfactory. ConclusionA good correlation was obtained among the standard, samples of polyherbal formulation and fractionated extract of O. gratissimum using HPTLC method.

Determination of ursolic acid of Sambucus adnata

HPLC was used to determine the content of ursolic acid of Sambucus adnata from different origins. The content of ursolic acid range between 1.14 to 5.7 microg, r = 0.999 8, the recovery range from 99.8% to 101.3%. The method is quick, sensitive and repeatable for determination of the content of ursolic acid of S. adnata.

Analysis of two anti-tumor active ingredients in Radix Actinidiae chinensis by dispersive liquid–liquid microextraction coupled to high performance liquid chromatography–mass spectrometry

A very simple and effective method for the extraction and determination of oleanolic acid (OA) and ursolic acid (UA) in Radix Actinidiae chinensis was developed using ultrasound-assisted dispersive liquid–liquid microextraction (UA-DLLME) and high performance liquid chromatography–mass spectrometry (HPLC–MS). Ethanol was used as the solvent to extract OA and UA from Radix Actinidiae chinensis, assisted by ultrasonic waves. When the extraction process was finished, the two target analytes in the extraction solvent were rapidly transferred from the ethanol extract to another extraction solvent (chloroform) with the help of a dispersive solvent (tetrahydrofuran) by using DLLME. A series of extraction parameters were investigated and optimized. Under the most favorable conditions, the limits of detection (LOD, S/N = 3) and enrichment factors were 2.0 ng g−1, 89.4 and 6.0 ng g−1, 84.5 for OA and UA, respectively. The recoveries of the spiked sample solutions at three different concentration levels of 200, 500 and 1000 ng g−1 ranged between 88.2 and 96.6%. The proposed method was the first successfully applied for the extraction and determination of OA and UA in Radix Actinidiae chinensis.

Determination of ursolic acid and ursolic acid lactone in the leaves of Eucalyptus tereticornis by HPLC

A simple isocratic HPLC method has been developed for the simultaneous quantification of two bioactive triterpenes, ursolic acid and ursolic acid lactone in E. tereticornis leaves. Samples were analyzed on RP-18 (4.6 × 250 mm, 5 µm) column with methanol and water acidified to pH 3.5 with TFA (88:12) at 210 nm. The method was validated and applied for the simultaneous quantification of the individual triterpenes in E. tereticornis extract. The calibration curves were linear over a concentration range of 0.05-0.3 mg mL -1 (r = 0.999 and 0.998, respectively). The limits of detection and quantification were 0.190 and 0.644 µg for ursolic acid, and 0.176 and 0.587 µg for ursolic acid lactone, while the percentage recoveries were 97.32 and 96.23% for ursolic acid and ursolic acid lactone, respectively. This is the first report on the HPLC method of ursolic acid lactone with high precision and accuracy.

Utilization of Dispersive Liquid-Liquid Microextraction Coupled with HPLC-UV as a Sensitive and Efficient Method for the Extraction and Determination of Oleanolic Acid and Ursolic Acid in Chinese Medicinal Herbs

Isomeric triterpenic acids of oleanolic acid (OA) and ursolic acid (UA) both have very low ultraviolet absorption and always exist in the same plant, so the separation and simultaneous determination of them have been a difficult task. In this study, a sensitive method combining dispersive liquid-liquid microextraction (DLLME) with HPLC-UV was developed for the extraction and determination of OA and UA in traditional Chinese medicinal herbs (CMHs). Variables influencing DLLME such as type and volume of extraction solvent, volume of dispersive solvent, ionic strength, aqueous phase pH, extraction time, centrifugation speed and time, and sample volume were investigated and optimized. Under the optimum conditions, both OA and UA attained favorable extraction efficiencies with enrichment factors 1378 and 933, respectively. The linear dynamic ranges of 0.07 - 30.4 μg?mL–1 for OA and 0.08 - 33.6 μg?mL–1 for UA were obtained with square correlation coefficients of 0.9963. The detection limits of OA and UA were both 0.02 μg?mL–1. The method recoveries ranged between 88.2% - 116.2% for OA and 85.7% - 108.2% for UA with the RSDs (n = 5) lower than 8.6%. The proposed method was successfully applied to concentrate and simultaneously determine these two triterpenic acids in Hedyotis diffusa and Eriobotrya japonica samples.

Quantification of Bioactive Principles in Indian Traditional Herb Ocimum Sanctum Linn. (Holy Basil) Leaves by High Performance Liquid Chromatography

A simple, rapid, accurate and reliable HPLC method for the determination of rosmarinic acid (RA) and ursolic acid (UA) in the leaves of ethanol extract of Ocimum sanctum (EEOS) has been developed for the first time and validated. Several biological activities of EEOS have been attributed to the presence of phenolic compounds and terpenes in the crude extract. The separation system consisted of a Phenomenex C18 reversed-phase column, using phosphate buffer / acetonitrile (80:20) and acetonitrile / water / methanol (90:5:5) as mobile phase for the determination of RA and UA respectively. The flow rate was 1.0 ml min−1 and detection wavelength was set at 326 and 261 nm for RA and UA respectively. The recovery of the method was in the range of 93.85 to 95.77 % for RA and 92.76 to 97.40 % for UA, and all the compounds showed good linearity in a relatively wide concentration range. Using the optimized conditions, the quantity of RA and UA in EEOS was found to be 0.26 ± 0.01% and 0.40 ± 0.01% w/w, respectively. The method is simple, sensitive, reproducible and ideally suited for rapid routine analysis.

Determination of Ursolic Acid in Force Loquat Capsule by Ultrasonic Extraction and Ionic Liquid Based Reverse Dispersive LLME

A new microextraction technique termed reverse dispersive liquid–liquid microextraction (R-DLLME) was developed. An orthogonal array design was used to optimize ultrasonic extraction conditions. In this method, the appropriate ionic liquid was injected into the sample by pipette. The performance of R-DLLME was illustrated with the determination of ursolic acid in force loquat capsule sample by RP-LC-UV detection. The optimized method exhibits a good linearity (r = 0.9989) over the studied range (2.0–60.0 μg mL−1) for ursolic acid. The recovery was 95.9–105.5%. The R-DLLME method has the advantages of simplicity of operation, rapidity, low cost, high recovery and enrichment factor.

A spectrophotometric method for the quantitative determination of ursolic acid

A spectrophotometric method has been developed for the quantitative determination of ursolic acid in industrial cranberry wastes and in the technical preparation obtained from them.